Intracellular imaging of nanoparticles: is it an elemental mistake to believe what you see?
Intracellular imaging of nanoparticles: is it an elemental mistake to believe what you see?
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DOI：
10.1186/1743-8977-7-15
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发表时间：
2010-06-03
影响因子：
10
通讯作者：
Rothen-Rutishauser B
中科院分区：
文献类型：
--
作者：
Brandenberger C;Clift MJ;Vanhecke D;Mühlfeld C;Stone V;Gehr P;Rothen-Rutishauser B
为了了解纳米颗粒（<100 nm）如何与细胞系统相互作用，可能导致不良反应，重要的是能够在细胞内检测和定位它们。由于纳米颗粒的尺寸很小，透射电子显微镜（TEM）是用于观察细胞内纳米颗粒的合适技术，因为光学显微镜无法在单个颗粒水平上分辨它们。然而，TEM细胞样品中其他细胞和非细胞纳米尺寸结构的存在，其可能在尺寸、形态和电子密度上类似于NP，可能阻碍NP的精确细胞内鉴定。因此，建议进行元素分析以确认细胞内NP的存在。本研究强调了进行元素分析的必要性，特别是能量过滤TEM，以确认使用量子点（QD）为例的细胞内NP定位。最近，量子点由于其荧光特性而受到越来越多的关注，并且已经提出了可能的生物医学成像应用。然而，不能排除潜在的不良反应，一些研究指出细胞内颗粒定位和毒性作用之间的相关性。将J774.A1鼠巨噬细胞样细胞暴露于NH2聚乙烯（PEG）QD，并使用电子光谱成像（ESI）对推定的细胞内QD进行QD中存在的两种元素（硫和镉）的元素共定位分析。这两种元素都显示在一个单一的颗粒水平和量子点被证实位于细胞内囊泡。然而，ESI分析表明，并非所有的纳米尺寸的结构，最初确定为量子点，得到证实。这一观察结果强调了使用TEM研究细胞内NP定位时进行元素分析的必要性。
In order to understand how nanoparticles (NPs <100 nm) interact with cellular systems, potentially causing adverse effects, it is important to be able to detect and localize them within cells. Due to the small size of NPs, transmission electron microscopy (TEM) is an appropriate technique to use for visualizing NPs inside cells, since light microscopy fails to resolve them at a single particle level. However, the presence of other cellular and non-cellular nano-sized structures in TEM cell samples, which may resemble NPs in size, morphology and electron density, can obstruct the precise intracellular identification of NPs. Therefore, elemental analysis is recommended to confirm the presence of NPs inside the cell. The present study highlights the necessity to perform elemental analysis, specifically energy filtering TEM, to confirm intracellular NP localization using the example of quantum dots (QDs). Recently, QDs have gained increased attention due to their fluorescent characteristics, and possible applications for biomedical imaging have been suggested. Nevertheless, potential adverse effects cannot be excluded and some studies point to a correlation between intracellular particle localization and toxic effects. J774.A1 murine macrophage-like cells were exposed to NH2 polyethylene (PEG) QDs and elemental co-localization analysis of two elements present in the QDs (sulfur and cadmium) was performed on putative intracellular QDs with electron spectroscopic imaging (ESI). Both elements were shown on a single particle level and QDs were confirmed to be located inside intracellular vesicles. Nevertheless, ESI analysis showed that not all nano-sized structures, initially identified as QDs, were confirmed. This observation emphasizes the necessity to perform elemental analysis when investigating intracellular NP localization using TEM.
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影响因子：
10.4
作者：
Hardman R
通讯作者：
Hardman R
影响因子：
3.8
作者：
Clift, Martin J. D.;Rothen-Rutishauser, Barbara;Stone, Vicki
通讯作者：
Stone, Vicki
影响因子：
3.8
作者：
Zhang, Leshuai W.;Monteiro-Riviere, Nancy A.
通讯作者：
Monteiro-Riviere, Nancy A.
影响因子：
1
作者：
SIMON, GT
通讯作者：
SIMON, GT
影响因子：
4.7
作者：
Lovric, J;Bazzi, HS;Maysinger, D
通讯作者：
Maysinger, D